Molecular identification ofSarcocystisspp. helped to define the origin of green pythons (Morelia viridis) confiscated in Germany

Abstract

Sarcocystis spp. represent apicomplexan parasites. They usually have a heteroxenous life cycle. Around 200 species have been described, affecting a wide range of animals worldwide, including reptiles. In recent years, large numbers of reptiles have been imported into Europe as pets and, as a consequence, animal welfare and species protection issues emerged. A sample of pooled feces from four confiscated green pythons (Morelia viridis) containing Sarcocystis spp. sporocysts was investigated. These snakes were imported for the pet trade and declared as being captive-bred. Full length 18S rRNA genes were amplified, cloned into plasmids and sequenced. Two different Sarcocystis spp. sequences were identified and registered as Sarcocystis sp. from M. viridis in GenBank. Both showed a 95-97% sequence identity with the 18S rRNA gene of Sarcocystis singaporensis. Phylogenetic analysis positioned these sequences together with other Sarcocystis spp. from snakes and rodents as definitive and intermediate hosts (IH), respectively. Sequence data and also the results of clinical and parasitological examinations suggest that the snakes were definitive hosts for Sarcocystis spp. that circulate in wild IH. Thus, it seems unlikely that the infected snakes had been legally bred. Our research shows that information on the infection of snakes with Sarcocystis spp. may be used to assess compliance with regulations on the trade with wildlife species.

Diagnosis of gastrointestinal parasites in reptiles: comparison of two coprological methods

“…and to some extent heterakids or ascarids (in tortoises and chameleons) show high prevalences in captive reptiles [ 20 ] and may pose a serious health problem for these animals while digenean trematodes will require one or more intermediate hosts, mainly molluscs. In terraria this cycle will most often be interrupted so that captive animals normally are not exposed to re-infection [ 20 ],[ 25 ],[ 31 ],[ 32 ]. Thus it seems advisable to examine captive reptiles by means of CNF rather than the SAF-method.…”

“…Furthermore, the detection of parasites with complicated life cycles as part of forensic parasitology gives helpful evidence for government officials dealing with legal regulations of exotic animals [ 45 ]. Large numbers of reptiles are being imported to Europe every year, and often government officials are challenged by the question whether these animals were farmed as documented in the import licence papers or taken directly from the wild [ 31 ],[ 32 ]. One such wild-caught green python ( Morelia viridis ) involved in this study harboured Kapsulotaenia spp.…”

“…Samples of Group B were obtained in 2011 from reptiles (mainly green pythons/ Morelia viridis ) which had been recently imported from Indonesia and had been taken into custody by local authorities to clarify whether they really were captive-bred individuals as declared upon import. Based on results from faecal parasitological examination and supported by further evidence obtained by physical and other laboratory examinations, strong evidence could be provided that many of these animals were indeed wild-caught [ 31 ],[ 32 ]. Samples of Group C were collected during 2011 directly at the reptile owners' domiciles in Switzerland, as part of a Master thesis at the Vetsuisse Faculty of the Universitiy Bern with the preselection to be wild-caught animals.…”

Molecular characterization of Sarcocystis spp. in intestine mucosal scrapings and fecal samples of Pampas fox (Lycalopex gymnocercus)

“…Consensus sequences obtained were compared with others reported in GenBank by BLASTn analysis. Samples with inconclusive amplicon sequencing results were considered as potential mixed infections of different protozoan species [14]. Only assembled sequences with high quality chromatograms were submitted to GenBank (accessions numbers listed in Table 1).…”

“…but also from other related coccidian. Therefore, samples that had shown inconclusive results in sequencing were considered as putative mixed infections [3,14,18]. Regarding species identification, only sequences with a BLASTn identity≥99% were considered as species specific and the respective sequence entered into GenBank along with the respective species name.…”